TY - JOUR
T1 - Techniques for accurate protein identification in shotgun proteomic studies of human, mouse, bovine, and chicken lenses
AU - Wilmarth, Phillip A.
AU - Riviere, Michael A.
AU - David, Larry L.
PY - 2009/12
Y1 - 2009/12
N2 - Analysis of shotgun proteomics datasets requires techniques to distinguish correct peptide identifications from incorrect identifications, such as linear discriminant functions and target/decoy protein databases. We report an efficient, flexible proteomic analysis workflow pipeline that implements these techniques to control both peptide and protein false discovery rates.We demonstrate its performance by analyzing two-dimensional liquid chromatography separations of lens proteins from human, mouse, bovine, and chicken lenses. We compared the use of International Protein Index databases to UniProt databases and no-enzyme SEQUEST searches to tryptic searches. Sequences present in the International Protein Index databases allowed detection of several novel crystallins. An alternate start codon isoform of βA4 was found in human lens. The minor crystallin γN was detected for the first time in bovine and chicken lenses. Chicken γS was identified and is the first member of the γ-crystallin family observed in avian lenses.
AB - Analysis of shotgun proteomics datasets requires techniques to distinguish correct peptide identifications from incorrect identifications, such as linear discriminant functions and target/decoy protein databases. We report an efficient, flexible proteomic analysis workflow pipeline that implements these techniques to control both peptide and protein false discovery rates.We demonstrate its performance by analyzing two-dimensional liquid chromatography separations of lens proteins from human, mouse, bovine, and chicken lenses. We compared the use of International Protein Index databases to UniProt databases and no-enzyme SEQUEST searches to tryptic searches. Sequences present in the International Protein Index databases allowed detection of several novel crystallins. An alternate start codon isoform of βA4 was found in human lens. The minor crystallin γN was detected for the first time in bovine and chicken lenses. Chicken γS was identified and is the first member of the γ-crystallin family observed in avian lenses.
KW - Bioinformatics
KW - False discovery rates
KW - Lens crystallins
KW - Shotgun proteomics
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U2 - 10.1007/s12177-009-9042-6
DO - 10.1007/s12177-009-9042-6
M3 - Article
AN - SCOPUS:76849095652
VL - 2
SP - 223
EP - 234
JO - Journal of Ocular Biology, Diseases, and Informatics
JF - Journal of Ocular Biology, Diseases, and Informatics
SN - 1936-8437
IS - 4
ER -